Peripheral Vascular Disease (PVD) results from chronic loss of the oxygen supply to the lower limbs due to arterial blockage. Loss of oxygen to these tissues can result in pain, ulcers, infection and limb amputation in patients. PVD affects up to 12% of the population globally with increased risk in patients with diabetes, hypertension, advanced age and obesity. Current therapies to treat mild and severe disease, including pharmaceutical interventions and revascularization, do not cure the disease but rather treat symptoms, can have significant side effects and are not successful in a large subset of patients. Development of the next generation of PVD therapies have been predicated on the hypothesis that increasing collateral blood vessel formation through angiogenesis will increase blood flow to affected tissues, thereby alleviating both the symptoms and the underlying cause of PVD. MultiStem(R), an allogenic large scale expanded stem cell population derived from bone marrow, have been shown to secrete multiple angiogenic factors and have been used in preclinical and clinical trials to treat ischemic injury. In our Phase I proposal, our primary objective is to examine the use of a small molecule to enhance the angiogenic activity of MultiStem and increase the benefit in a hind limb ischemia model. To further develop the therapeutic potential of these cells, we propose to investigate increasing the angiogenic benefit of MultiStem by the use of a small molecule, DFO (Deferoxamine Mesylate). First, treatment of cells with a small molecule, DFO, a HIF11 inducer, will be optimized to maximize the angiogenic potential of the cells as determined in an in vivo matrigel plug assay. Hypoxia induces the secretion of angiogenic factors in many cell lines through the induction of the transcriptional regulator, HIF11. Preliminary studies have shown that treating the cells with a HIF11 inducer increases their secretion of angiogenic factors and we hypothesize this treatment will improve the therapeutic potential of the cells. Second, the effects MultiStem treated with or without DFO will be examined to determine whether modulation of MultiStem angiogenic potential by a small molecular provides a superior benefit in enhancing collateral function in a mouse hind limb ischemia model. Finally, the mechanism of action of treatment with MultiStem will be further examined by determining the extent of blood vessel formation and endogenous progenitor cell recruitment to the site of injury in tissues of hind limb ischemia model animals treated with MultiStem compared to vehicle treated controls. Upon successful completion of this study, we will transition to a Phase II pre-clinical disease model for safety and benefit testing in a GLP animal model that will be part of an IND package to the FDA. It is our intention to commercialize a stem cell therapeutic product for the treatment of peripheral vascular disease. PUBLIC HEALTH RELEVANCE: Relevance: use of a small molecule to enhance the efficacy and therapeutic benefit of stem cells for the treatment of peripheral vascular disease.